Hydrogel, inkjet ink, method for producing cell culture body, and cell-containing gel particles and method for producing same

Abstract

An object of the present invention is to provide a hydrogel that has both gel shapeability and cell adhesiveness, and enables a long-time cell culture. Provided is a hydrogel ( 1 ) including: a tetra-armed polymer ( 2 ) containing polyethylene glycol as a backbone, and containing one or more electrophilic functional groups or nucleophilic functional groups in at least one of a side chain and a terminal; and a linear polymer ( 3 ) containing two or more nucleophilic functional groups or electrophilic functional groups in at least one of a side chain and a terminal; the tetra-armed polymer ( 2 ) and the linear polymer ( 3 ) being cross-linked with each other; wherein the hydrogel is characterized in that the linear polymer ( 3 ) is protein, peptide, or polysaccharide, that the electrophilic functional group(s) is/are one or more selected from the group consisting of maleimidyl, N-hydroxy-succinimidyl (NHS), sulfosuccinimidyl, phthalimidyl, imidazoyl, acryloyl, and nitrophenyl, and that the nucleophilic functional group(s) is/are one or more selected from the group consisting of thiol, amino, and —CO 2 PhNO 2 .

1 . A hydrogel, comprising:
 a multibranched polymer containing polyethylene glycol as a backbone, and containing one or more electrophilic functional groups or nucleophilic functional groups in at least one of a side chain and a terminal; and   a linear polymer containing two or more nucleophilic functional groups or electrophilic functional groups in at least one of a side chain and a terminal;   the multibranched polymer and the linear polymer being cross-linked with each other, wherein   the linear polymer is protein, peptide, or polysaccharide,   the electrophilic functional group(s) is/are one or more selected from the group consisting of maleimidyl, N-hydroxy-succinimidyl (NHS), sulfosuccinimidyl, phthalimidyl, imidazoyl, acryloyl, and nitrophenyl, and   the nucleophilic functional group(s) is/are one or more selected from the group consisting of thiol, amino, and —CO 2 PhNO 2 .   
     
     
         2 . The hydrogel according to  claim 1 , wherein the linear polymer is gelatin. 
     
     
         3 . The hydrogel according to  claim 1 , further comprising at least one cell. 
     
     
         4 . The hydrogel according to  claim 3 , wherein the at least one cell comprises at least two or more species of cells. 
     
     
         5 . The hydrogel according to  claim 3 , wherein the cell is a neurocyte. 
     
     
         6 . An ink jet ink comprising:
 a first solution comprising a multibranched polymer containing polyethylene glycol as a backbone, and containing one or more electrophilic functional groups or nucleophilic functional groups in at least one of a side chain and a terminal; and   a second solution including a linear polymer containing two or more nucleophilic functional groups or electrophilic functional groups in at least one of a side chain and a terminal, wherein   the linear polymer is protein, peptide, or polysaccharide,   the electrophilic functional group(s) is/are one or more selected from the group consisting of maleimidyl, N-hydroxy-succinimidyl (NHS), sulfosuccinimidyl, phthalimidyl, imidazoyl, acryloyl, and nitrophenyl, and   the nucleophilic functional group(s) is/are one or more selected from the group consisting of thiol, amino, and —CO 2 PhNO 2 .   
     
     
         7 . The ink jet ink according to  claim 6 , wherein the linear polymer is gelatin. 
     
     
         8 . The ink jet ink according to  claim 6 , wherein one or both of the first solution and the second solution comprises a cell. 
     
     
         9 . The ink jet ink according to  claim 8 , wherein the cell is a neurocyte. 
     
     
         10 . A method for producing a cell culture body, comprising:
 retaining a first solution including a multibranched polymer containing polyethylene glycol as a backbone, and having one or more functional groups in at least one of a side chain and a terminal, any of the functional group(s) being one of a nucleophilic functional group and an electrophilic functional group; and   landing droplets of a second solution on the first solution to form one or more hydrogels, the droplets being discharged from a droplet-discharging device in such a manner that the second solution comes into contact with the first solution, the second solution including a linear polymer having one or more functional groups in at least one of a side chain and a terminal, any of the functional group(s) being the other one of the nucleophilic functional group and the electrophilic functional group, wherein   the linear polymer is protein, peptide, or polysaccharide,   the electrophilic functional group(s) is/are one or more selected from the group consisting of maleimidyl, N-hydroxy-succinimidyl (NHS), sulfosuccinimidyl, phthalimidyl, imidazoyl, acryloyl, and nitrophenyl,   the nucleophilic functional group(s) is/are one or more selected from the group consisting of thiol, amino, and —CO 2 PhNO 2 , and   at least one of the first solution and the second solution contains a cell.   
     
     
         11 . The method for producing a cell culture body according to  claim 10 , wherein, when the first solution and the second solution are brought into contact with each other, at least one of the first solution and the second solution is discharged by an ink jet method. 
     
     
         12 . The method for producing a cell culture body according to  claim 11 , wherein the first solution and the second solution are brought into contact with each other at a resolution of 500 μm or less. 
     
     
         13 . A cell-containing gel particle comprising a hydrogel and a cell encapsulated in the hydrogel, the hydrogel comprising:
 a multibranched polymer containing polyethylene glycol as a backbone, and containing one or more electrophilic functional groups or nucleophilic functional groups in at least one of a side chain and a terminal; and   a linear polymer containing two or more nucleophilic functional groups or electrophilic functional groups in at least one of a side chain and a terminal;   the multibranched polymer and the linear polymer being cross-linked with each other, wherein   the linear polymer is protein, peptide, or polysaccharide,   the electrophilic functional group(s) is/are one or more selected from the group consisting of maleimidyl, N-hydroxy-succinimidyl (NHS), sulfosuccinimidyl, phthalimidyl, imidazoyl, acryloyl, and nitrophenyl, and   the nucleophilic functional group(s) is/are one or more selected from the group consisting of thiol, amino, and —CO 2 PhNO 2 .   
     
     
         14 . The cell-containing gel particle according to  claim 13 , wherein the linear polymer is gelatin. 
     
     
         15 . The cell-containing gel particle according to  claim 13 , having a diameter of from 60 μm to 500 μm. 
     
     
         16 . A method for producing a cell-containing gel particle, comprising mixing the following:
 a first solution including a multibranched polymer containing polyethylene glycol as a backbone, and containing one or more electrophilic functional groups or nucleophilic functional groups in at least one of a side chain and a terminal; and   a second solution including a linear polymer containing two or more nucleophilic functional groups or electrophilic functional groups in at least one of a side chain and a terminal, wherein   the linear polymer is protein, peptide, or polysaccharide,   the electrophilic functional group(s) is/are one or more selected from the group consisting of maleimidyl, N-hydroxy-succinimidyl (NHS), sulfosuccinimidyl, phthalimidyl, imidazoyl, acryloyl, and nitrophenyl,   the nucleophilic functional group(s) is/are one or more selected from the group consisting of thiol, amino, and —CO 2 PhNO 2 ,   one of the first solution and the second solution contains a cell, and   a droplet of one which is the first solution or the second solution, and contains the cell is mixed with the other one which is the first solution or the second solution, and does not contain the cell.   
     
     
         17 . The method for producing a cell-containing gel particle according to  claim 16 , wherein the linear polymer is gelatin. 
     
     
         18 . A method for culturing a cell, comprising: placing the cell-containing gel particle according to  claim 13  in a culture container; and culturing a cell in such a manner that the cell-containing gel particle is not in contact with the culture container. 
     
     
         19 . A method for culturing a cell, comprising: allowing the cell-containing gel particle according to  claim 13  to adhere to a culture container; and culturing a cell in such a manner that the cell-containing gel particle is retained in the culture container. 
     
     
         20 . A cell-evaluating plate comprising the cell-containing gel particle according to  claim 13 .